Effect and Mechanism of Kir2.1 Channel Overexpression on Transdifferentiation of Endothelial Progenitor Cells

Q4 Biochemistry, Genetics and Molecular Biology Molecular & Cellular Biomechanics Pub Date : 2019-02-21 DOI:10.32604/MCB.2019.05753
Jifeng Li, Yanting He, Xiaoyun Zhang, Hong Li, Xiumei Guan, Min Cheng, Xiaodong Cui
{"title":"Effect and Mechanism of Kir2.1 Channel Overexpression on Transdifferentiation of Endothelial Progenitor Cells","authors":"Jifeng Li, Yanting He, Xiaoyun Zhang, Hong Li, Xiumei Guan, Min Cheng, Xiaodong Cui","doi":"10.32604/MCB.2019.05753","DOIUrl":null,"url":null,"abstract":"Objective The propose of the study is to investigate the specific effects of the mechanically sensitive channel Kir2.1 on the transdifferentiation of EPCs so as to understand the molecular mechanism of pathological vascular remodeling. Methods Endothelial progenitor cells (EPCs) were isolated from rat bone marrow and cultured in EGM2 medium in vitro. The recombinant lentiviral vectors carrying Kir2.1 (NM_017296.1) gene was designed and constructed in order to overexpress the gene. The smooth muscle cells (SMCs) molecules marker on EPCs, such as α-SMA, FSP1 and α-SM22, were detected by RT-PCR and cellular immunofluorescence. In addition, cell angiogenic capacity and migration in vitro were assessed by Matrigel and Transwell methods respectively. Moreover, neointimal thickening was evaluated in the surgery model of balloon injury of rat carotid artery in vivo. Result The results showed that the expression levels of α-SM22, FSP1 and α-SMA were up-regulated in the Kir2.1 overexpression group compared with the control. The number of migrating cells in the Kir2.1 overexpression group was significantly higher than that in the scramble group, while quantitative assessment further confirmed that the Kir2.1 overexpression strongly attenuated the ability of bone marrow-derived EPC to form tube-like structures in Matrigel assay. Compared with the control group, morphometric analysis showed ratio of intimal area/medial area (I/M) in rats was increased in rats transplanted with Lenti-Kir2.1 overexpression. Conclusion It is indicated that the overexpression of channel Kir2.1 induces EPCs transdifferentiated into mesenchymal transition SMCs (EndoMT). It may provide a potential target for the treatment or prevention of pathological vascular remodeling disease.","PeriodicalId":48719,"journal":{"name":"Molecular & Cellular Biomechanics","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2019-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular & Cellular Biomechanics","FirstCategoryId":"1087","ListUrlMain":"https://doi.org/10.32604/MCB.2019.05753","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 1

Abstract

Objective The propose of the study is to investigate the specific effects of the mechanically sensitive channel Kir2.1 on the transdifferentiation of EPCs so as to understand the molecular mechanism of pathological vascular remodeling. Methods Endothelial progenitor cells (EPCs) were isolated from rat bone marrow and cultured in EGM2 medium in vitro. The recombinant lentiviral vectors carrying Kir2.1 (NM_017296.1) gene was designed and constructed in order to overexpress the gene. The smooth muscle cells (SMCs) molecules marker on EPCs, such as α-SMA, FSP1 and α-SM22, were detected by RT-PCR and cellular immunofluorescence. In addition, cell angiogenic capacity and migration in vitro were assessed by Matrigel and Transwell methods respectively. Moreover, neointimal thickening was evaluated in the surgery model of balloon injury of rat carotid artery in vivo. Result The results showed that the expression levels of α-SM22, FSP1 and α-SMA were up-regulated in the Kir2.1 overexpression group compared with the control. The number of migrating cells in the Kir2.1 overexpression group was significantly higher than that in the scramble group, while quantitative assessment further confirmed that the Kir2.1 overexpression strongly attenuated the ability of bone marrow-derived EPC to form tube-like structures in Matrigel assay. Compared with the control group, morphometric analysis showed ratio of intimal area/medial area (I/M) in rats was increased in rats transplanted with Lenti-Kir2.1 overexpression. Conclusion It is indicated that the overexpression of channel Kir2.1 induces EPCs transdifferentiated into mesenchymal transition SMCs (EndoMT). It may provide a potential target for the treatment or prevention of pathological vascular remodeling disease.
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Kir2.1通道过表达对内皮祖细胞转分化的影响及机制
目的探讨机械敏感通道Kir2.1在EPCs转分化中的具体作用,以了解病理性血管重构的分子机制。方法从大鼠骨髓中分离内皮祖细胞(EPCs),体外培养于EGM2培养基中。设计并构建了携带Kir2.1 (NM_017296.1)基因的重组慢病毒载体,以过表达该基因。采用RT-PCR和细胞免疫荧光法检测EPCs上平滑肌细胞(SMCs)分子标志物α-SMA、FSP1和α-SM22。此外,分别采用Matrigel法和Transwell法评估细胞的体外血管生成能力和迁移能力。并在大鼠颈动脉球囊损伤手术模型中对其新生内膜增厚进行了评价。结果Kir2.1过表达组与对照组相比,α-SM22、FSP1和α-SMA的表达水平均上调。Kir2.1过表达组的迁移细胞数量明显高于scramble组,同时在Matrigel实验中进一步定量评估证实,Kir2.1过表达强烈减弱了骨髓源性EPC形成管状结构的能力。形态计量学分析显示,与对照组相比,lentii - kir2.1过表达移植大鼠的内膜面积/内侧面积(I/M)比增加。结论过表达Kir2.1通道可诱导EPCs向间充质过渡型SMCs (EndoMT)转化。它可能为治疗或预防病理性血管重构疾病提供一个潜在的靶点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 去求助
来源期刊
Molecular & Cellular Biomechanics
Molecular & Cellular Biomechanics CELL BIOLOGYENGINEERING, BIOMEDICAL&-ENGINEERING, BIOMEDICAL
CiteScore
1.70
自引率
0.00%
发文量
21
期刊介绍: The field of biomechanics concerns with motion, deformation, and forces in biological systems. With the explosive progress in molecular biology, genomic engineering, bioimaging, and nanotechnology, there will be an ever-increasing generation of knowledge and information concerning the mechanobiology of genes, proteins, cells, tissues, and organs. Such information will bring new diagnostic tools, new therapeutic approaches, and new knowledge on ourselves and our interactions with our environment. It becomes apparent that biomechanics focusing on molecules, cells as well as tissues and organs is an important aspect of modern biomedical sciences. The aims of this journal are to facilitate the studies of the mechanics of biomolecules (including proteins, genes, cytoskeletons, etc.), cells (and their interactions with extracellular matrix), tissues and organs, the development of relevant advanced mathematical methods, and the discovery of biological secrets. As science concerns only with relative truth, we seek ideas that are state-of-the-art, which may be controversial, but stimulate and promote new ideas, new techniques, and new applications.
期刊最新文献
Hot Topics of Molecular and Cellular Biomechanics in 2022 CFD Study on Hemodynamic Characteristics of Inferior Vena Cava Filter Affected by Blood Vessel Diameter Can PAPE-Induced Increases in Jump Height Be Explained by Jumping Kinematics? Reconstruction of the Hindlimb Locomotion of Confuciusornis (Aves) and Its Implication for the Origin of Avian Flight Classification of Leukemia and Leukemoid Using VGG-16 Convolutional Neural Network Architecture
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1